Method and composition for detecting phosphorylating compounds



" 'METHOD COMPOSITION FOR DETECTING PHOSPHORYLATING COMPOUNDS .David N. Kramer, Baltimore, 'Md.,-assigno'rto the United States of America, as represented ,by the. Secretary of theArmy Serial No. 621,183

10 Claims. 3 (Cl."23230) (Granted under Title 35, U. S. Code (1952), sec. 266) "No'Dra wing. 'pplication November 8, 1956 The invention described hereinmaybe manufactured ,and, used ,by. or. for the. Government :of the United States also known as GB, and related compounds, with oximes and to compositions for detecting thepresence of such compounds by. the use of that reaction.

We have found thatphosphorylating compoundsyielding acid anions on hydrolysis will react with substituted monoximes of alpha, beta dicarbonyl compounds and that when the reaction is carried out in the presence of Congo red, a red to blue color develops. The reaction is also accompanied by frothing. Other acid-base indicators operative in the pH range 3.5-5 may be substituted for Congo red. In each case the reaction causes a change to the acid color of the indicator.

Example 1 A drop of acid-free isopropyl methylphosphonofluoridate was brought into contact with a butanedione monoxime-Congo red mixture. The color changed from red to blue. Frothing was also observed in the reaction mixture. Congo red in the absence of butanedione monoxime was found to be unaifected by the isopropyl methylphosphonofluoridate.

Example 2 A series of oximes was then screened for reactivity towards isopropyl methylphospho-nofiuoridate in the following manner. A mixture of 0.1 g. oxime and 0.05 g. Congo red was. dispersed in 0.5 g. blanc fixe and the pale red powder was placed in beakers. A drop of acid-free isopropyl methylphosphonofiuoridate was then placed on compounds:

Butanedlone monoxime CHaC-C-CH:

N 03 and 2-phenylglyoxal-1-mon0xime O-C-CH 2,865,719 Patented Dec. 23, 1958 A, fast reaction, indicated by a bluejcolor accompanied by framing, was obtained with 2-methylglyoxal-l-monoxime CH3-C'CH l-phenylglyoxnl-l-monoxime-O UE lion ,8

Negative results were obtainedwith the-simple oximes, alpha hydroxy oximesand the dioximes tested. Compounds whichrgaveanegative result were, acetone oxime, acetaldoxime, ben zaldoxirne, --salicy-laldoxime, benzoinoxime, and dimethylglyoxime, among-others. v

The reaction described herein is ,to;be f'distinguished from-that of Fischer, Mille -Sass and W-itten in .{application-S. N. 621,184, filed of-even date herewith. In. the latter case, diisonitrosoacetone (acetonedioxirne), is-employed as a detector for various compounds, including isopropyl methylphosphonofiuoridate, under alkaline conditions. In my procedure, I'employ alpha carbonyl monoximes together'with an :acid-base indicator operative in the pH 3.5-5 range. v

In addition to isopropyl -methylphosphone-fluoridate, this procedure may be employedtoidetect other phosphorlyating compounds-yielding acid anions on hydrolysis, for example: pinacolyl methylphosphonofiuori'date, also known-as-GD,

and

'0 CHs-F CH:

-CH-CCHa CH3 CH3 cyclohexyi methylphosphonofluoridate, also known as GF 0 CHr-I F GHQ-CH:

CH CH2 \CH2 (32 $12 1 dimethylphosphoramide cyanidate, also known as 0 (CH:)2Nl-CN -C2Hs and diisopropyltluorophosphine oxide, also known as DFP CH; O

As a test procedure, a mixture of an alpha keto oxime, an acid base indicator and an inert diluent, which maybe blanc fixe (barium sulfate) or other material which will not neutralize the liberated acid, e. g. HF, is exposed to liquids or vapors to be tested. A change of the indicator to the acid color indicates the presence of the compounds identified above. The mixture may be made up in the form of a chalk.

I believe the mechanism of the reaction to be as follows, using GB as the reactant.

*To explainthe observation that frothing occurs simul-- taneously with liberation of HF in this reaction, it is postulated that the phosphorylated form of the alpha ketook'im (1) decomposes in the presence of the acid to yield gas, e. g., N and the hydrolysis products of GB and the= alpha k'eto o'xime'. Y

While I have given certain specific examples, it will be: obvious that various changes are possible. I therefore-v wish my invention to be limited solely by the scope of the: appended claims.

I claim:

1. A composition of matter for detecting the presence of a phosphorylating compound yielding acid anions on hydrolysis and selected from the group consisting of isopropyl methylphosphonofiuoridate, pinacolyl 1nethylphos phonofluoridate, cyclohexyl rnethylphosphonofluoridate, ethyl dimethylphosphoroamidocyarnidate, and diisopropylfluorophosphine oxide comprising a mixture of an alpha carbonyl monoxime, an acid-base indicator showing a change in the pH 3.5-5 range, and an inert diluent, said alpha carbonyl monoxime having the formula wherein R and R are selected from the group consisting of methyl, phenyl and hydrogen.

2. A composition as defined in claim 1 wherein said oxime is butanedione monoxime.

3. A composition as defined in claim 1 wherein said oxime is Z-methylglyoxal-l-monoxime.

4. A composition as defined in claim 1 wherein said oxime is 2-phenylglyoxal-l-monoxime.

S. A composition as defined in claim I wherein said oxime is l-phenylglyoxal-l-monoxime.

6. The method detecting a phosphorylating compound yielding acid anions on hydrolysis and selected from the group consisting of isopropyl methylphosphonofluoridate, pinacolyl methylphosphonofiuoridate, cyclohexyl methylphospho'nofiuoridate, ethyl dimethylphosphoroamidocyamidate, and diisopropylfluorophcsphine oxide which comprises reacting said compound with an alpha carbonyl monoxime in the presence of an acid-base indicator-showing a color change in the pH range 3.5 to 5, said alpha carbonyl monoxime havingthe formula References Cited in the file of this patent UNITED STATES PATENTS Dieterich Jan. 14, 1902 Truog Nov. 17,1931 

1. A COMPOSITION OF MATTER FOR DETECTING THE PRESENCE OF A PHOSPHORYLATING COMPOUND YIELDING ACID ANIONS ON HYDROLYSIS AND SELECTED FROM THE GROUP CONSISTING OF ISOPROPYL METHYLPHOSPHONOFLUORIDATE, PINCOLYL METHYLPHOSPHONOFLUORIDATE, CYCLOHEXYL METHYLPHOSPHONOFLUORIDATE, ETHYL DIMETHYLPHOSPHOROAMIDACYAMIDATE, AND DIISOPROPYLFLUOROPHOSPHINE OXIDE COMPRISING A MIXTURE OF AN ALPHA CARBONYL MONOXIME, AN ACID-BASE INDICATOR SHOWING A CHANGE IN THE PH 3.5-5 RANGE, AND AN INERT DILUENT, SAID ALPHA CARBONYL MONOXIME HAVING THE FORMULA 